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Effects of Connexin 32-Mediated Lung Inflammation Resolution During Liver Ischemia Reperfusion

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Abstract

Background

Hepatic ischemia reperfusion (HIR) leads to a lung inflammatory response and subsequent pulmonary barrier dysfunction. The gap junction communication protein connexin 32 (Cx32), which is widely expressed in the lungs, participates in intercellular signaling. This study determined whether the communication protein Cx32 could affect pulmonary inflammation caused by HIR.

Methods

Mice were randomly allocated into four groups (n = 8/group): (i) Cx32+/+ sham group; (ii) Cx32+/+ HIR model group; (iii) Cx32−/− sham group; and (iv) Cx32−/− HIR model group. Twenty-four hours after surgery, lung tissues were collected for bright field microscopy, western blot (Cx32, JAK2, p-JAK2, STAT3, p-STAT3), and immunofluorescence (ZO-1, 8-OHDG) analyses. The collected bronchoalveolar fluid was tested for levels of interleukin-6 (IL-6), matrix metalloproteinase 12 (MMP-12), and antitrypsin (α1-AT). Lung mmu-miR-26a/b expression was detected using a PCR assay.

Results

Increased expression of Cx32 mRNA and protein was noted in the lungs after HIR. Cx32 deletion significantly aggravated pulmonary function from acute lung injury induced by HIR. In addition, Cx32 deletion decreased the protein level of ZO-1 (pulmonary function) and increased the level of the oxidative stress marker 8-OHDG in the lungs. Moreover, in the Cx32−/− HIR model group, the levels of IL-6 and MMP-12 in bronchoalveolar lavage fluid were significantly increased leading to activation of the JAK2/STAT3 pathway, and decreased α1-AT levels. Furthermore, we found mmu-miR-26a/b was significantly downregulated in the Cx32−/− HIR model group.

Conclusion

HIR leads to acute lung inflammatory injury. Cx32 deletion aggravates hepatic-derived lung inflammation, partly through blocking the transferring of mmu-miR-26a/b and leading to IL-6-related JAK2/STAT3 pathway activation.

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References

  1. Douzinas EE, Livaditi O, Tasoulis MK, et al. Nitrosative and oxidative stresses contribute to post-ischemic liver injury following severe hemorrhagic shock: the role of hypoxemic resuscitation. PLoS ONE. 2012;7:e32968.

    Article  CAS  Google Scholar 

  2. Darnis B, Mohkam K, Schmitt Z, et al. Subtotal hepatectomy in swine for studying small-for-size syndrome and portal inflow modulation: is it reliable? HPB (Oxford). 2015;17:881–888.

    Article  Google Scholar 

  3. Madadi-Sanjani O, Kuebler JF, Dippel S, et al. Long-term outcome and necessity of liver transplantation in infants with biliary atresia are independent of cytokine milieu in native liver and serum. Cytokine. 2018;111:382–388.

    Article  CAS  Google Scholar 

  4. Ulger BV, Erbis H, Turkcu G, et al. Nebivolol ameliorates hepatic ischemia/reperfusion injury on liver but not on distant organs. J Invest Surg. 2015;28:245–252.

    Article  Google Scholar 

  5. Mikrou A, Kalimeris KA, Lilis I, et al. Molecular studies of the immunological effects of the sevoflurane preconditioning in the liver and lung in a rat model of liver ischemia/reperfusion injury. Mol Immunol. 2016;72:1–8.

    Article  CAS  Google Scholar 

  6. Lepere V, Vanier A, Loncar Y, et al. Risk factors for pulmonary complications after hepatic resection: role of intraoperative hemodynamic instability and hepatic ischemia. BMC Anesthesiol. 2017;17:84.

    Article  Google Scholar 

  7. Li GZ, Sloane JL, Lidsky ME, et al. Simultaneous diaphragm and liver resection: a propensity-matched analysis of postoperative morbidity. J Am Coll Surg. 2013;216:402–411.

    Article  Google Scholar 

  8. Li X, Wei X, Chen C, et al. N-Acetylcysteine inhalation improves pulmonary function in patients received liver transplantation. Biosci Rep. 2018;38:pii: BSR20180858.

    Article  Google Scholar 

  9. Ge M, Chen C, Yao W, et al. Overexpression of Brg1 alleviates hepatic ischemia/reperfusion-induced acute lung injury through antioxidative stress effects. Oxid Med Cell Longev. 2017;2017:8787392.

    Article  CAS  Google Scholar 

  10. Sperber J, Lipcsey M, Larsson A, et al. Evaluating the effects of protective ventilation on organ-specific cytokine production in porcine experimental postoperative sepsis. BMC Pulm Med. 2015;15:60.

    Article  CAS  Google Scholar 

  11. Coulon P, Landisman CE. The potential role of gap junctional plasticity in the regulation of state. Neuron. 2017;93:1275–1295.

    Article  CAS  Google Scholar 

  12. Freund-Michel V, Muller B, Marthan R, et al. Expression and role of connexin-based gap junctions in pulmonary inflammatory diseases. Pharmacol Ther. 2016;164:105–119.

    Article  CAS  Google Scholar 

  13. Losa D, Chanson M, Crespin S. Connexins as therapeutic targets in lung disease. Expert Opin Ther Targets. 2011;15:989–1002.

    Article  CAS  Google Scholar 

  14. Okamoto T, Akiyama M, Takeda M, et al. Connexin32 is expressed in vascular endothelial cells and participates in gap-junction intercellular communication. Biochem Biophys Res Commun. 2009;382:264–268.

    Article  CAS  Google Scholar 

  15. Okamoto T, Akiyama M, Takeda M, et al. Connexin32 protects against vascular inflammation by modulating inflammatory cytokine expression by endothelial cells. Exp Cell Res. 2011;317:348–355.

    Article  CAS  Google Scholar 

  16. Zhang Y, Yuan D, Yao W, et al. Hyperglycemia aggravates hepatic ischemia reperfusion injury by inducing chronic oxidative stress and inflammation. Oxid Med Cell Longev. 2016;2016:3919627.

    PubMed  PubMed Central  Google Scholar 

  17. Yao W, Li H, Luo G, et al. SERPINB1 ameliorates acute lung injury in liver transplantation through ERK1/2-mediated STAT3-dependent HO-1 induction. Free Radic Biol Med. 2017;108:542–553.

    Article  CAS  Google Scholar 

  18. He Y, Ye ZQ, Li X, et al. Alpha7 nicotinic acetylcholine receptor activation attenuated intestine-derived acute lung injury. J Surg Res. 2016;201:258–265.

    Article  CAS  Google Scholar 

  19. Miyajima A, Ohashi H, Fujishiro A, et al. Effects of all trans-retinoic acid on alveolar regeneration in dexamethasone-induced emphysema models and its relationship to exposure in ICR and FVB mice. Biol Pharm Bull. 2016;39:927–934.

    Article  CAS  Google Scholar 

  20. Li H, Yao W, Liu Z, et al. Hyperglycemia abrogates ischemic postconditioning cardioprotection by impairing AdipoR1/Caveolin-3/STAT3 signaling in diabetic rats. Diabetes. 2015;65:942–955.

    Article  CAS  Google Scholar 

  21. Zhao W, Zhou S, Yao W, et al. Propofol prevents lung injury after intestinal ischemia-reperfusion by inhibiting the interaction between mast cell activation and oxidative stress. Life Sci. 2014;108:80–87.

    Article  CAS  Google Scholar 

  22. Chi X, Yao W, Zhang A, et al. Downregulation of lung toll-like receptor 4 could effectively attenuate liver transplantation-induced pulmonary damage at the early stage of reperfusion. Mediators Inflamm. 2015;2015:383907.

    Article  CAS  Google Scholar 

  23. Yao W, Luo G, Zhu G, et al. Propofol activation of the Nrf2 pathway is associated with amelioration of acute lung injury in a rat liver transplantation model. Oxid Med Cell Longev. 2014;2014:258567.

    Article  CAS  Google Scholar 

  24. Luo G, Zhu G, Yuan D, et al. Propofol alleviates acute lung injury following orthotopic autologous liver transplantation in rats via inhibition of the NADPH oxidase pathway. Mol Med Rep. 2015;11:2348–2354.

    Article  CAS  Google Scholar 

  25. Luo GJ, Yao WF, He Y, et al. Ulinastatin prevents acute lung injury led by liver transplantation. J Surg Res. 2015;193:841–848.

    Article  CAS  Google Scholar 

  26. Colletti LM, Kunkel SL, Walz A, et al. Chemokine expression during hepatic ischemia/reperfusion-induced lung injury in the rat. The role of epithelial neutrophil activating protein. J Clin Invest. 1995;95:134–141.

    Article  CAS  Google Scholar 

  27. Nastos C, Kalimeris K, Papoutsidakis N, et al. Global consequences of liver ischemia/reperfusion injury. Oxid Med Cell Longev. 2014;2014:906965.

    Article  CAS  Google Scholar 

  28. Yu Z, Tong Y, Zhang R, et al. Saquinavir ameliorates liver warm ischemia-reperfusion-induced lung injury via HMGB-1- and P38/JNK-Mediated TLR-4-dependent signaling pathways. Mediators Inflamm. 2017;2017:7083528.

    PubMed  PubMed Central  Google Scholar 

  29. Schulz R, Gorge PM, Gorbe A, et al. Connexin 43 is an emerging therapeutic target in ischemia/reperfusion injury, cardioprotection and neuroprotection. Pharmacol Ther. 2015;153:90–106.

    Article  CAS  Google Scholar 

  30. O’Donnell JJ III, Birukova AA, Beyer EC, et al. Gap junction protein connexin43 exacerbates lung vascular permeability. PLoS One. 2014;9:e100931.

    Article  Google Scholar 

  31. Rignault S, Haefliger JA, Waeber B, et al. Acute inflammation decreases the expression of connexin 40 in mouse lung. Shock. 2007;28:78–85.

    Article  CAS  Google Scholar 

  32. Duffy HS, Iacobas I, Hotchkiss K, et al. The gap junction protein connexin32 interacts with the Src homology 3/hook domain of discs large homolog 1. J Biol Chem. 2007;282:9789–9796.

    Article  CAS  Google Scholar 

  33. Proboszcz M, Paplinska-Goryca M, Nejman-Gryz P, et al. A comparative study of sTREM-1, IL-6 and IL-13 concentration in bronchoalveolar lavage fluid in asthma and COPD: a preliminary study. Adv Clin Exp Med. 2017;26:231–236.

    Article  Google Scholar 

  34. Ruwanpura SM, Mcleod L, Miller A, et al. Interleukin-6 promotes pulmonary emphysema associated with apoptosis in mice. Am J Respir Cell Mol Biol. 2011;45:720–730.

    Article  CAS  Google Scholar 

  35. Chen M, Liu Y, Yi D, et al. Tanshinone IIA promotes pulmonary artery smooth muscle cell apoptosis in vitro by inhibiting the JAK2/STAT3 signaling pathway. Cell Physiol Biochem. 2014;33:1130–1138.

    Article  CAS  Google Scholar 

  36. Fujimoto E, Sato H, Shirai S, et al. Connexin32 as a tumor suppressor gene in a metastatic renal cell carcinoma cell line. Oncogene. 2005;24:3684–3690.

    Article  CAS  Google Scholar 

  37. Hong X, Sin WC, Harris AL, et al. Gap junctions modulate glioma invasion by direct transfer of microRNA. Oncotarget. 2015;6:15566–15577.

    Article  Google Scholar 

  38. Yang X, Liang L, Zhang XF, et al. MicroRNA-26a suppresses tumor growth and metastasis of human hepatocellular carcinoma by targeting interleukin-6-Stat3 pathway. Hepatology. 2013;58:158–170.

    Article  CAS  Google Scholar 

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Funding

The study was supported in part by grants from Natural Science Foundation of China (No.81974081 and No.81601724 for Weifeng Yao; No.81974296 and No.81772127 and No. 81571926 for Ziqing Hei; No.81770649 for Gangjian Luo) and the Science and Technology Project of Guangdong Province (No. 2019A1515011852 to Weifeng Yao) the Young Teacher Training Program of Sun Yat-sen University, China (Grant no. 19ykpy23 for Weifeng Yao).

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Author notes

  1. Zheng Zhang and Weifeng Yao have contributed equally to this work.

Authors and Affiliations

  1. Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510630, China

    Zheng Zhang, Weifeng Yao, Dongdong Yuan, Fei Huang, Yue Liu, Gangjian Luo & Ziqing Hei

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  1. Zheng Zhang
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  2. Weifeng Yao
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  3. Dongdong Yuan
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Contributions

Zheng Zhang and Weifeng Yao participated in research design, data collection, and data analysis and wrote the manuscript. Ziqing Hei and Gangjian Luo participated in research design, data collection, and data analysis. Dongdong Yuan, Fei Huang, Yue Liu participated in data collection and data analysis. All authors participated meaningfully in the study and have seen and approved the final manuscript.

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Correspondence to Ziqing Hei.

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Zhang, Z., Yao, W., Yuan, D. et al. Effects of Connexin 32-Mediated Lung Inflammation Resolution During Liver Ischemia Reperfusion. Dig Dis Sci 65, 2914–2924 (2020). https://doi.org/10.1007/s10620-019-06020-8

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